Abstract
The mechanical state of cells is a critical part of their healthy functioning and it is controlled primarily by cytoskeletal networks (actin, microtubules and intermediate filaments). Drug-based strategies targeting the assembly of a given cytoskeletal network are often used to pinpoint their role in cellular function. Unlike actin and microtubules, there has been limited interest in the role of intermediate filaments, and fewer drugs have thus been identified and characterised as modulators of its assembly. Here, we evaluate whether Withaferin-A (WFA), an established disruptor of vimentin filaments, can also be used to modulate keratin filament assembly. Our results show that in keratinocytes, which are keratin-rich but vimentin-absent, Withaferin-A disrupts keratin filaments. Importantly, the dosages required are similar to those previously reported to disrupt vimentin in other cell types. Furthermore, Withaferin-A-induced keratin disassembly is accompanied by changes in cell stiffness and migration. Therefore, we propose that WFA can be repurposed as a useful drug to disrupt the keratin cytoskeleton in epithelial cells.
Highlights
Drug-based disruption of cytoskeletal (CSK) networks offers a rapid, dose-responsive and often reversible technique for studying mechanotransduction and cell signalling
We show that WFA can be of similar use in keratin-rich cell types, as previously shown in vimentin-rich cell types, opening up its re-purposing for studying Keratin filaments and expanding the arsenal of cyto-modulatory drugs
To visualise the Human Epidermal Keratinocytes (HEKs) cells and their cytoskeleton, cells were stained for Keratin 14 (K14), α-tubulin and actin simultaneously
Summary
Drug-based disruption of cytoskeletal (CSK) networks offers a rapid, dose-responsive and often reversible technique for studying mechanotransduction and cell signalling. IFs have emerged as important players in cells’ mechanical properties: they impart protection on the nucleus during migration [1] and are capable of withstanding high strains [2] They are biomarkers in certain types of cancer [3], and when mutated or altered in terms of expression or assembly, can lead to a unique set of diseases with aberrant tissue mechanics [4,5]. One such drug is Withaferin-A (WFA), which is used to disrupt vimentin It was put forth by Bargagna-Mohan [11] and is derived from the ayurvedic medicine Ashwagandha. We report that in Human Epidermal Keratinocytes (HEKs), WFA disrupts keratin assembly at similar concentrations as those used for Vimentin disruption in other cell types. We show that WFA can be of similar use in keratin-rich cell types, as previously shown in vimentin-rich cell types, opening up its re-purposing for studying Keratin filaments and expanding the arsenal of cyto-modulatory drugs
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